Methods for detecting and modulating the sensitivity of tumor cells to anti-mitotic agents and for modulating tumorigenicity

Abstract

Provided herein is a method of screening a tumour cell for resistance to a tubulin-binding agent, the method comprising detecting the expression of any one or more of class II, class III and class IVb β-tubulin by the tumour cell, wherein the expression of any one or more of class II, class III and class IVb β-tubulin indicates that the tumour cell has resistance or potential resistance to the tubulin-binding agent. Also provided are methods of modulating the sensitivity of a tumour cell Also provided is a method of modulating the tumorigenesis of a tumour cell.

What is claimed is: 
     
         1 . A method of screening a tumour cell for resistance to a tubulin-binding agent, the method comprising detecting expression of any one or more of class II, class III and class IVb β-tubulin by the tumour cell, wherein the expression of any one or more of class II, class III and class IVb β-tubulin indicates that the tumour cell has resistance or potential resistance to the tubulin-binding agent. 
     
     
         2 . The method according to  claim 1  wherein detecting expression of any one or more of class II, class III and class IVb β-tubulin by the tumour cell comprises detecting the expression of any two of class II, class III and class IVb β-tubulin by the tumour cell. 
     
     
         3 . The method according to  claim 1 , wherein detecting expression of any one or more of class II, class III and class IVb β-tubulin by the tumour cell comprises detecting the expression of class II, class III and class IVb β-tubulin by the tumour cell. 
     
     
         4 . The method according to  claim 1 , wherein detecting expression of one or more of class II, class III and class IVb β-tubulin by the tumour cell comprises detecting the expression of any one or more of class II, class III and class IVb β-tubulin protein by the tumour cell. 
     
     
         5 . The method according to  claim 1 , wherein detecting expression of any one or more of class II, class III and class IVb β-tubulin by the tumour cell comprises detecting the expression of any one or more of class II, class III and class IVb β-tubulin mRNA by the tumour cell. 
     
     
         6 . The method according to  claim 1 , wherein detecting expression of one or more of class II, class III and class IVb β-tubulin by the tumour cell comprises quantifying the level of expression of any one or more of class II, class III and class IVb β-tubulin by the tumour cell. 
     
     
         7 . The method according to  claim 1 , wherein detecting expression of one or more of class II, class III and class IVb β-tubulin by the tumour cell comprises comparing the expression of one or more of class II, class III and class IVb β-tubulin by the tumour cell with the expression of one or more of class II, class III and class IVb β-tubulin by a control cell. 
     
     
         8 . A method of assessing resistance of a tumour cell to a tubulin-binding agent, the method comprising:
 detecting an amount of any one or more of class II, class III and class IVb β-tubulin protein or any one or more of class II, class III and class IVb β-tubulin mRNA by the tumour cell and thereby obtaining a subject tumour cell tubulin amount,   comparing the subject tumour cell tubulin amount to a control cell tubulin amount thereby determining a tubulin amount difference, wherein the control cell tubulin amount is a respective amount of a class II, class III or class IVb β-tubulin protein or any one or more of class II, class III and class IVb β-tubulin mRNA in a control cell, and   assessing resistance of the tumour cell to the tubulin-binding agent based on the tubulin amount difference.   
     
     
         9 . The method according to  claim 8 , wherein the control cell is a control non tumour cell. 
     
     
         10 . The method according to  claim 8 , wherein the control cell is a control tumour cell. 
     
     
         11 . The method according to  8 , wherein the control tumour cell is a tumour cell which is resistant to the tubulin-binding agent. 
     
     
         12 . The method according to  claim 8 , wherein the control tumour cell is a tumour cell which is sensitive to the tubulin-binding agent. 
     
     
         13 . The method according to  claim 8 , wherein detecting an amount of any one or more of class II, class III and class IVb β-tubulin protein is detecting an amount of class II, class III and class IVb β-tubulin protein. 
     
     
         14 . The method according to  claim 8 , wherein detecting an amount of any one or more of class II, class III and class IVb β-tubulin protein is detecting an amount of class III β-tubulin protein only. 
     
     
         15 . A method of screening a tumour cell for resistance to a DNA damaging agent, the method comprising detecting the expression of class III β-tubulin by the tumour cell, wherein the expression of class III β-tubulin indicates that the tumour cell has resistance or potential resistance to the DNA damaging agent. 
     
     
         16 . A method for enhancing sensitivity of a tumour cell to a tubulin-binding agent, the method comprising introducing into the tumour cell an effective amount of at least one nucleic acid construct comprising a nucleotide sequence specific to at least a portion of the class II, class III or class IVb β-tubulin gene product, wherein the construct decreases the expression of class II, class III or class IVb β-tubulin in the tumour cell and thereby increases the sensitivity of the tumour to the tubulin-binding agent. 
     
     
         17 . A method for enhancing sensitivity of a tumour cell to a DNA damaging agent, the method comprising introducing into the tumour cell an effective amount of at least one nucleic acid construct comprising a nucleotide sequence specific to at least a portion of the class III β-tubulin gene product, wherein the construct decreases the expression of class III β-tubulin in the tumour cell and thereby increases the sensitivity of the tumour to the DNA damaging agent. 
     
     
         18 . A method for treating a tumour cell in a subject, comprising administering to the subject an effective amount of at least one nucleic acid construct comprising a nucleotide sequence specific to at least a portion of the class II, class III or class IVb β-tubulin gene product and administering to the subject a tubulin-binding agent, wherein the nucleic acid construct decreases the expression of class II, class III or class IVb β-tubulin in the tumour and thereby increases the sensitivity of the tumour cell to the tubulin-binding agent. 
     
     
         19 . A method for treating a tumour cell in a subject, comprising administering to the subject an effective amount of at least one nucleic acid construct comprising a nucleotide sequence specific to at least a portion of the class III β-tubulin gene product and administering to the subject a DNA damaging agent, wherein the nucleic acid construct decreases the expression of class III β-tubulin in the tumour and thereby increases the sensitivity of the tumour cell to the DNA damaging agent. 
     
     
         20 . The method of  claim 18 , comprising administering to the subject a tubulin binding agent in addition to the DNA damaging agent. 
     
     
         21 . The method according to  claim 18 , wherein the nucleic acid construct and the tubulin-binding agent are administered simultaneously. 
     
     
         22 . The method according to  claim 18 , wherein the nucleic acid construct and the tubulin-binding agent are administered concurrently. 
     
     
         23 . The method according to  claim 18 , wherein the nucleic acid construct is administered before the tubulin-binding agent. 
     
     
         24 . The method according to  claim 18  wherein the nucleotide sequence is selected from the group consisting of an antisense sequence, an siRNA sequence, a shRNA sequence and a ribozyme sequence. 
     
     
         25 . The method according to  claim 18 , wherein the nucleotide sequence is specific to at least a portion of the class IVb β-tubulin gene product. 
     
     
         26 . The method according to  claim 18 , wherein the nucleotide sequence is specific to at least a portion of the class II β-tubulin gene product. 
     
     
         27 . The method according to  claim 18 , wherein the nucleotide sequence is specific to at least a portion of the class III β-tubulin gene product. 
     
     
         28 . The method according to  claim 18 , wherein the tubulin-binding agent is a microtubule destabilizing agent. 
     
     
         29 . The method according to  claim 18 , wherein the tubulin-binding agent is a microtubule destabilizing agent which is selected from the group consisting of any a vinca alkaloid, a dolostatin, a colchicine, a cryptophycin, curacin A, and 2-methoxyestradiol. 
     
     
         30 . The method according to  claim 18 , wherein the tubulin-binding agent is a microtubule stabilizing agent. 
     
     
         31 . The method according to  claim 18 , wherein the microtubule stabilizing agent is a taxane or an epothilone. 
     
     
         32 . The method according to  claim 19  wherein the DNA damaging agent is selected from the group consisting of adriamycin, bleomycin, etoposide or a platinum compound. 
     
     
         33 . The method according to  claim 19  wherein the DNA damaging agent is selected from the group consisting of cisplatin, trans-analogue of cisplatin, carboplatin, oxaliplatin, nedaplatin, iproplatin, and tetraplatin. 
     
     
         34 . The method according to  claim 1 , wherein the tumour cell is a non-small cell lung carcinoma cell. 
     
     
         35 . The method according to  claim 18 , wherein the tumour cell is a non-small cell lung carcinoma cell. 
     
     
         36 . A pharmaceutical composition for increasing sensitivity of a tumour to a tubulin-binding agent, the pharmaceutical composition comprising at least one nucleic acid construct comprising a nucleotide sequence specific to at least a portion of the class II, class III or class IVb β-tubulin gene, wherein the construct is able to decrease the expression of class II, class III or class IVb β-tubulin in a tumour, together with a pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         37 . A pharmaceutical composition for increasing sensitivity of a tumour to a DNA damaging agent, the pharmaceutical composition comprising at least one nucleic acid construct comprising a nucleotide sequence specific to at least a portion of the class III β-tubulin gene, wherein the construct is able to decrease the expression of class III β-tubulin in a tumour, together with a pharmaceutically acceptable carrier, diluent or excipient 
     
     
         38 . A kit for assessing sensitivity of a tumour to a tubulin-binding agent, the kit comprising at least one nucleic acid construct comprising a nucleotide sequence specific to at least a portion of the class II, class III or class IVb β-tubulin gene product, or at least one antibody specific to at least one of the class II, class III or class IVb β-tubulin protein, wherein the construct or antibody is used to detect the expression of class II, class III or class IVb β-tubulin in the tumour. 
     
     
         39 . The kit according to  claim 38 , further comprising a tubulin-binding agent. 
     
     
         40 . A kit for assessing sensitivity of a tumour to a DNA damaging agent, the kit comprising at least one nucleic acid construct comprising a nucleotide sequence specific to at least a portion of the class III β-tubulin gene product, or at least one antibody specific to class III β-tubulin protein, wherein the construct or antibody is used to detect the expression of class III β-tubulin in the tumour. 
     
     
         41 . A method of reducing tumorigenicity of a class III β-tubulin expressing tumour cell in a subject, the method comprising administering to the subject a nucleic acid molecule comprising a polynucleotide sequence which is specific to at least a portion of the class III β-tubulin gene, wherein the polynucleotide sequence is able to decrease the expression of class III β-tubulin in the tumour cell and thereby reduce the tumorigenicity of the tumour cell. 
     
     
         42 . A pharmaceutical composition for reducing tumorigenicity of a class III β-tubulin expressing tumour cell in a subject, the pharmaceutical composition comprising a nucleic acid molecule which comprises a polynucleotide sequence which is specific to at least a portion of the class III β-tubulin gene, wherein the polynucleotide sequence is able to decrease the expression of class III β-tubulin in the tumour cell, together with a pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         43 . The composition according to  claim 42 , wherein the polynucleotide sequence is selected from an antisense sequence, an siRNA sequence, a ribozyme sequence, a microRNA sequence and an shRNA sequence. 
     
     
         44 . The method according to  claim 41 , wherein the polynucleotide sequence is selected from an antisense sequence, an siRNA sequence, a ribozyme sequence, a microRNA sequence and an shRNA sequence. 
     
     
         45 . The method according to  claim 41 , wherein the class III β-tubulin expressing tumour cell is a non-small cell lung carcinoma cell (NSCLC), an ovarian cancer cell, a breast cancer cell, a gastric cancer cell, a melanoma cell, a prostate cancer cell, or a cell from a tumour of unknown origin. 
     
     
         46 . The composition according to  claim 42 , wherein the class III β-tubulin expressing tumour cell is a non-small cell lung carcinoma cell (NSCLC), an ovarian cancer cell, a breast cancer cell, a gastric cancer cell, a melanoma cell, a prostate cancer cell, or a cell from a tumour of unknown origin.